Modulation of cellular cholesterol by MβCD:MβCD-cholesterol mixtures in Bovine Aortic Endothelial cells (A) and in Chinese Hamster Ovary Cells (B). Modified from Romanenko et al. (2002) [74] and Byfield et al., [135]. MβCD saturated with cholesterol using the standard protocol. Briefly, small volume of a cholesterol stock solution in chloroform:methanol (1:1, vol:vol) was added to a glass tube and the solvent was evaporated. Then, 2.5 mM or 5 mM MβCD solution in DMEM medium without serum was added to the dried cholesterol. The tube was vortexed, sonicated, and incubated overnight in a shaking bath at 37°C. The mixture of MβCD and MβCD -cholesterol was prepared just by mixing the two solutions at different molar ratios.